Separation of Nicotinamide and Related Substances on Zodiac HST TH Column

A complex mixture containing nicotinamide and its related impurities was successfully separated using the Zodiac HST TH mixed-mode column. Compounds such as nicotinamide, methylnicotinamide, nicotinamide adenine mononucleotide (NMN), nicotinamide adenine dinucleotide (NAD), and adenosine monophosphate (AMP) were baseline resolved within a 15-minute method.

The Zodiac HST TH column employs a trimodal separation mechanism—reversed-phase, cation-exchange, and anion-exchange—which enables high selectivity for structurally similar and highly polar compounds. Retention and resolution can be finely tuned by adjusting the acetonitrile content, buffer concentration, and pH. Additional selectivity is achievable by exploring different buffer systems within the same pH range. This approach is ideal for the analysis of nucleotides and related biomolecules in complex mixtures.